Biochemical Journal最新文献_第2页

Identification of aSinorhizobium meliloti YbgC-like thioesterase that contributes to the production of the infochemical 2-tridecanone. 促进信息化学物质2-三烯酮生成的亚硝酸根菌ybgc样硫酯酶的鉴定。

IF 4.1 3区生物学

Biochemical Journal Pub Date : 2025-09-13 DOI: 10.1042/bcj20253120

Lydia M Bernabéu-Roda,Geovanny Rivera-Hernández,Virginia Cuéllar,Rafael Núñez,Ángeles Moreno-Ocampo,Christian Sohlenkamp,Otto Geiger,María J Soto,Isabel M López-Lara

{"title":"Identification of aSinorhizobium meliloti YbgC-like thioesterase that contributes to the production of the infochemical 2-tridecanone.","authors":"Lydia M Bernabéu-Roda,Geovanny Rivera-Hernández,Virginia Cuéllar,Rafael Núñez,Ángeles Moreno-Ocampo,Christian Sohlenkamp,Otto Geiger,María J Soto,Isabel M López-Lara","doi":"10.1042/bcj20253120","DOIUrl":"https://doi.org/10.1042/bcj20253120","url":null,"abstract":"Sinorhizobium meliloti is a soil bacterium that can establish beneficial symbiosis with legume plants. The fadD gene encodes a long-chain fatty acyl-coenzyme A (CoA) synthetase. Inactivation of FadD in S. meliloti leads to a pleiotropic phenotype, including the overproduction of several volatile methylketones (MKs). One of them, 2-tridecanone (2-TDC), was found to act as an infochemical that affects important bacterial traits and hampers plant-bacteria interactions. Knowledge about bacterial genes involved in MK production is limited. In wild tomato species, MK synthesis requires intermediates of fatty acid biosynthesis and the activity of the methylketone synthase 2 (MKS2), a thioesterase belonging to the hot dog-fold family. In this study, we have identified SMc03960, a conserved hypothetical protein with homology to bacterial YbgC-like thioesterases, as an ortholog of MKS2 in S. meliloti. Heterologous expression of smc03960 in Escherichia coli results in the formation of several MKs, including 2-TDC, and causes the accumulation of free fatty acids. Purified His-SMc03960 showed thioesterase activity for different acyl groups linked either to acyl carrier protein (ACP) or to CoA with preference for C14-long substrates. Moreover, formation of 2-TDC in vitro was achieved by using His-SMc03960 and 3-oxo-myristoyl-ACP. Although deletion of smc03960 in the wild type or in the fadD mutant does not significantly alter the amount of MKs released by S. meliloti, overexpression of the gene results in increased production of 2-TDC in these two strains. Overall, our data demonstrate that SMc03960 is an acyl-ACP/acyl-CoA thioesterase with broad substrate specificity that contributes to 2-TDC formation.","PeriodicalId":8825,"journal":{"name":"Biochemical Journal","volume":"33 1","pages":""},"PeriodicalIF":4.1,"publicationDate":"2025-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145036037","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

RNMT-dependent RNA cap methylation in health and disease. rnmt依赖性RNA帽甲基化在健康和疾病中的作用。

IF 4.1 3区生物学

Biochemical Journal Pub Date : 2025-09-09 DOI: 10.1042/bcj20253170

Jacquie G Mills,Lydia A Hepburn,Victoria H Cowling

引用次数: 0

GSK-3α regulates miRNAs associated with transcriptional and metabolic processes in human cardiomyocytes under hypoxia. GSK-3α在缺氧条件下调控与人心肌细胞转录和代谢过程相关的mirna。

IF 4.3 3区生物学

Biochemical Journal Pub Date : 2025-09-09 DOI: 10.1042/BCJ20253208

Firdos Ahmad, Hezlin Marzook, Musa Idris, Omama I Dawuod, Megna Srinivas, Asima Karim, Mohamed A Saleh, Rizwan Qaisar

{"title":"GSK-3α regulates miRNAs associated with transcriptional and metabolic processes in human cardiomyocytes under hypoxia.","authors":"Firdos Ahmad, Hezlin Marzook, Musa Idris, Omama I Dawuod, Megna Srinivas, Asima Karim, Mohamed A Saleh, Rizwan Qaisar","doi":"10.1042/BCJ20253208","DOIUrl":"10.1042/BCJ20253208","url":null,"abstract":"Glycogen synthase kinase-3α (GSK-3α) is a multifunctional kinase that plays roles in the pathogenesis of various cardiac diseases, including ischemia and pressure overload and ischemia-reperfusion-induced injury. It regulates key cellular processes such as cardiac cell proliferation, apoptosis, metabolism, and inflammation. However, its role in regulating cardiac microRNAs (miRNAs) remains unknown. To explore the role of GSK-3α in regulating miRNAs, we conducted an unbiased miRNA sequencing analysis in human GSK-3α-overexpressing AC16 cardiomyocytes (GOCs) under hypoxic conditions. Transcriptomic analysis demonstrated numerous differentially expressed miRNAs (DEMs) crucial for transcriptional, inflammatory, and various metabolic processes in the cell. Among 184 DEMs, hsa-miR-3934-5p, hsa-miR-139-5p, and hsa-miR-185-5p were the most up-regulated, while hsa-miR-193b-3p, hsa-miR-181a-2-3p, and hsa-miR-369-3p were the most down-regulated in GOC vs. control cells subjected to hypoxia. Gene ontology (GO) term analysis demonstrated a significant set of genes associated with the terms regulation of transcription, cellular protein modification process, cellular aromatic compound metabolic process, and nucleotide binding in GOC. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis further revealed enrichment of key pathways including metabolic, cytokine-cytokine receptor interaction, cyclic adenosine monophosphate (cAMP), and mitogen-activated protein kinase (MAPK) signaling pathways in GOC challenged with hypoxia. Collectively, these findings reveal a novel mechanism by which GSK-3α regulates a network of miRNAs in human cardiomyocytes required for critical transcriptional, metabolic, and signaling responses including the MAPK and inflammatory pathways under hypoxic stress. GSK-3α-mediated miRNA dysregulation may contribute to the pathophysiological changes observed in ischemia-induced cardiac injury.","PeriodicalId":8825,"journal":{"name":"Biochemical Journal","volume":" ","pages":""},"PeriodicalIF":4.3,"publicationDate":"2025-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144940743","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Mechanistic insights into DNA binding by BD1 of the TAF1 tandem bromodomain module. TAF1串联溴结构域模块BD1结合DNA的机制。

IF 4.1 3区生物学

Biochemical Journal Pub Date : 2025-09-08 DOI: 10.1042/bcj20253064

Yogita Yadav,Phibarisha Chyne,Babu Sudhamalla

{"title":"Mechanistic insights into DNA binding by BD1 of the TAF1 tandem bromodomain module.","authors":"Yogita Yadav,Phibarisha Chyne,Babu Sudhamalla","doi":"10.1042/bcj20253064","DOIUrl":"https://doi.org/10.1042/bcj20253064","url":null,"abstract":"Transcription initiation factor TFIID subunit 1 (TAF1) is a pivotal component of the TFIID complex, critical for RNA polymerase II-mediated transcription initiation. However, the molecular basis by which TAF1 recognizes and associates with chromatin remains incompletely understood. Here, we report that the tandem bromodomain module of TAF1 engages nucleosomal DNA through a distinct positively charged surface patch on the first bromodomain (BD1). Electrostatic potential mapping and molecular docking revealed a prominent basic region on BD1 that facilitates interaction with DNA, predominantly driven by hydrogen bonds and electrostatic forces, as supported by molecular dynamics simulations. Site-directed mutagenesis identified three key positively charged residues (R1435, K1436, and R1437) within the αA helix of BD1, constituting an \"RKR\" basic patch essential for DNA binding. Electrophoretic mobility shift assays demonstrated that the TAF1 tandem bromodomain binds DNA in a concentration-dependent manner with moderate preference for AT-rich sequences, attributed to this RKR motif. Importantly, DNA binding occurs independently of histone acetyllysine recognition by the bromodomains, as acetylated histone H4 peptides or mutations in the acetyllysine-binding pocket did not affect DNA interaction. Furthermore, nucleosome pulldown assays revealed that disruption of the BD1 RKR patch significantly reduces binding to acetylated nucleosomes, highlighting its role in facilitating chromatin engagement. Collectively, our findings establish the RKR basic patch on TAF1 BD1 as a critical determinant for DNA interaction, providing mechanistic insight into how TAF1 tandem bromodomains coordinate dual recognition of nucleosomal DNA and histone acetylation. These results offer a molecular basis for understanding how TAF1 may contribute to transcriptional regulation via chromatin engagement.","PeriodicalId":8825,"journal":{"name":"Biochemical Journal","volume":"28 1","pages":""},"PeriodicalIF":4.1,"publicationDate":"2025-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145025621","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The ClpA chaperone and the two adaptor proteins modulate the fate of the model substrate tagged with a SsrA-degron of Leptospira. ClpA伴侣和两个接头蛋白调节了钩端螺旋体SsrA-degron标记的模型底物的命运。

IF 4.3 3区生物学

Biochemical Journal Pub Date : 2025-09-04 DOI: 10.1042/BCJ20253143

Surbhi Kumari, Manish Kumar

{"title":"The ClpA chaperone and the two adaptor proteins modulate the fate of the model substrate tagged with a SsrA-degron of Leptospira.","authors":"Surbhi Kumari, Manish Kumar","doi":"10.1042/BCJ20253143","DOIUrl":"10.1042/BCJ20253143","url":null,"abstract":"Bacterial caseinolytic protease (Clp) chaperone-protease complexes are essential for the degradation of misfolded and aggregated protein substrates. The spirochaete Leptospira interrogans possesses a set of Clp adaptor proteins (ClpS1 and ClpS2) and chaperones (ClpX, ClpA and ClpC), which are believed to associate with two distinct isoforms of ClpP (ClpP1 and ClpP2). This study explores the structural and functional properties of LinClpA, LinClpS1 and LinClpS2 derived from L. interrogans. LinClpA, a 740-amino acid protein, features an N-terminal domain and two AAA+ ATPase domains (D-I and D-II), containing conserved motifs critical for ATP binding and hydrolysis. LinClpS1 and LinClpS2 exhibit similar structures, yet they possess distinct binding pockets for N-degron substrates. Biochemical assays indicate that the N-domain-deleted variant of LinClpA (LinClpAΔN) exhibits a nucleotide-induced oligomerization tendency similar to LinClpA's but demonstrates higher ATPase activity. Interaction studies have shown that LinClpA's ATPase activity is enhanced in the presence of LinClpP isoforms and inhibited by LinClpS isoforms. In contrast, the activity of LinClpAΔN remained unaffected by LinClpS1 and LinClpS2, highlighting the significance of the N-domain of LinClpA in adaptor protein interactions. Furthermore, the study predicted and evaluated the role of the C-degron tag called small stable RNA A in facilitating protein degradation by the L. interrogans ClpAP1P2 machinery.","PeriodicalId":8825,"journal":{"name":"Biochemical Journal","volume":" ","pages":"1253-1275"},"PeriodicalIF":4.3,"publicationDate":"2025-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12493166/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144940765","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

JMJD6 and YBX1 physically interact and regulate HOTAIR proximal promoter. JMJD6和YBX1物理相互作用并调控HOTAIR近端启动子。

IF 4.3 3区生物学

Biochemical Journal Pub Date : 2025-09-04 DOI: 10.1042/BCJ20243020

Aritra Gupta, Siddharth Bhardwaj, Kartiki V Desai

{"title":"JMJD6 and YBX1 physically interact and regulate HOTAIR proximal promoter.","authors":"Aritra Gupta, Siddharth Bhardwaj, Kartiki V Desai","doi":"10.1042/BCJ20243020","DOIUrl":"10.1042/BCJ20243020","url":null,"abstract":"Earlier, we showed that jumonji domain containing protein 6 (JMJD6) interacted with HOTAIR promoter (-123 to -103 bp, termed JMJD6 interaction region [JIR]) and for maximal induction, an additional (-216 to -123 bp) region was required. In silico prediction and ENCODE data from MCF7 cells showed Y-box interacting protein 1 (YBX1) peaks in this region (YIR). Publicly available mass spectrometry data of proteins following JMJD6 immunoprecipitation identified YBX1 as an interacting partner. In this study, we validate JMJD6-YBX1 interaction in breast cancer cell lines using co-immunoprecipitation assays with recombinant, endogenous and in vitro synthesized proteins. Domain mapping using deletion constructs revealed that the A/P domain of YBX1 interacted with the JMJC domain of JMJD6. These proteins also positively regulated each other's expression in breast cancer cell lines. Further, YBX1 augmented luciferase activity of HOTAIR promoter constructs, pHP216 and pHP123, in MCF7, Vec and JMJD6 overexpressing cells. siRNA-mediated depletion, mutation of YIR region or knocking out YBX1 (YKO cells) diminished luciferase activity. ChIP and ChIP-re-ChIP assays verified co-occupancy of both proteins in the HOTAIR promoter region. Electrophoretic mobility shift assays confirmed complex formation with YIR and JIR probes. Mutation of the YIR region and YKO resulted in loss of complex formation with both probes. Taken together, these data imply that YBX1 is crucial for physically recruiting JMJD6 to the HOTAIR promoter. Their interaction and positive feed-forward loop, perpetuated by JMJD6 and YBX1 inter-regulation, culminates in HOTAIR induction, which in turn is known to drive tumour progression.","PeriodicalId":8825,"journal":{"name":"Biochemical Journal","volume":" ","pages":"1289-1305"},"PeriodicalIF":4.3,"publicationDate":"2025-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144940748","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Selectivity profiles and substrate recognition of Rab-phosphorylating kinases. rabb磷酸化激酶的选择性和底物识别。

IF 4.3 3区生物学

Biochemical Journal Pub Date : 2025-09-04 DOI: 10.1042/BCJ20253212

Deep Chatterjee, Verena Dederer, Landon Vu Nguyen, Marcel Wendel, Kamal R Abdul Azeez, Swetha Mahesula, Florian Stengel, Samara Reck-Peterson, Sebastian Mathea

{"title":"Selectivity profiles and substrate recognition of Rab-phosphorylating kinases.","authors":"Deep Chatterjee, Verena Dederer, Landon Vu Nguyen, Marcel Wendel, Kamal R Abdul Azeez, Swetha Mahesula, Florian Stengel, Samara Reck-Peterson, Sebastian Mathea","doi":"10.1042/BCJ20253212","DOIUrl":"https://doi.org/10.1042/BCJ20253212","url":null,"abstract":"The Rab GTPase switch-2 region is a hotspot for post-translational modifications. Its phosphorylation can determine whether individuals develop Parkinson's disease or not. Other modifications of the same region are catalyzed by enzymes from bacterial pathogens when they infect human cells. Here, we profiled a set of kinases including LRRK1, LRRK2, DYRK1A, MST1 and TBK1 for their capability of phosphorylating Rab GTPases. We identified several novel kinase:Rab pairs, such as LRRK1:Rab43 and TBK1:Rab29. Further, we comprehensively assessed what makes a Rab GTPase a good kinase substrate, considering the Rab nucleotide-binding state and the Rab primary sequence. In a systematic mutational study, Rab variants with modulated phosphorylation properties were established, leading to the identification of a LRRK2 recognition patch in the Rab α3 helix. A Glu to Arg exchange in that patch increased the phosphorylation 18-fold, indicating that Rabs are suboptimal LRRK2 substrates. Given that this effect is also observed in a cellular model, we propose that our variants will be excellent tools for analysing the physiological function of Rab phosphorylation.","PeriodicalId":8825,"journal":{"name":"Biochemical Journal","volume":"482 17","pages":""},"PeriodicalIF":4.3,"publicationDate":"2025-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144991337","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Itaconate utilisation by the human pathogen Pseudomonas aeruginosa requires uptake via the IctPQM TRAP transporter. 人类病原体铜绿假单胞菌利用衣康酸需要通过IctPQM TRAP转运体摄取。

IF 4.3 3区生物学

Biochemical Journal Pub Date : 2025-08-28 DOI: 10.1042/BCJ20253132

Javeria Mehboob, Reyme Herman, Rory C Elston, Heritage Afolabi, Bethan E Kinniment-Williams, Marjan W van der Woude, Anthony J Wilkinson, Gavin H Thomas

{"title":"Itaconate utilisation by the human pathogen Pseudomonas aeruginosa requires uptake via the IctPQM TRAP transporter.","authors":"Javeria Mehboob, Reyme Herman, Rory C Elston, Heritage Afolabi, Bethan E Kinniment-Williams, Marjan W van der Woude, Anthony J Wilkinson, Gavin H Thomas","doi":"10.1042/BCJ20253132","DOIUrl":"https://doi.org/10.1042/BCJ20253132","url":null,"abstract":"Pseudomonas aeruginosa PA01 is one of the major causes of disease persistence and mortality in patients with lung pathologies, relying on various host metabolites as carbon and energy sources for growth. The ict-ich-ccl operon (pa0878, pa0882 and pa0883) in PAO1 is required for growth on the host molecule itaconate, a C5-dicarboxylate. However, it is not known how itaconate is taken up into P. aeruginosa. Here, we demonstrate that a genetically linked tripartite ATP-independent periplasmic (TRAP) transporter (pa0884-pa0886), which is homologous to the known C4-dicarboxylate-binding TRAP system, is essential for growth on itaconate, but not for the closely related C4-dicarboxylate succinate. Using tryptophan fluorescence spectroscopy, we demonstrate that the substrate-binding protein (SBP), IctP (PA0884), binds itaconate but still retains higher affinity for the related C4-dicarboxylates. The structures of IctP bound to itaconate (1.80 Å) and succinate (1.75 Å) revealed an enclosed ligand-binding pocket with ion pairing interactions with the ligand carboxylates. The C2 methylene group that is the distinguishing feature of itaconate compared with succinate is accommodated by a unique change in the IctP-binding site from a Leu to Val, which distinguishes it from closely related C4-dicarboxylate-binding SBPs. Together, these data suggest that this transporter, which we name IctPQM, has duplicated from a canonical C4-dicarboxylate transporter, and its evolution towards itaconate specificity enables this pathogen to now access a key metabolite for persistence in the host.","PeriodicalId":8825,"journal":{"name":"Biochemical Journal","volume":"482 17","pages":""},"PeriodicalIF":4.3,"publicationDate":"2025-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144940722","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Allosteric regulation of proteolytic machines unveiled by the synergy between cryo-EM and solution NMR spectroscopy. 低温电镜和溶液核磁共振波谱的协同作用揭示了蛋白水解机器的变构调节。

IF 4.3 3区生物学

Biochemical Journal Pub Date : 2025-08-26 DOI: 10.1042/BCJ20253239

Madison Turner, Robert W Harkness, Zev A Ripstein, Rui Huang, Siavash Vahidi

{"title":"Allosteric regulation of proteolytic machines unveiled by the synergy between cryo-EM and solution NMR spectroscopy.","authors":"Madison Turner, Robert W Harkness, Zev A Ripstein, Rui Huang, Siavash Vahidi","doi":"10.1042/BCJ20253239","DOIUrl":"10.1042/BCJ20253239","url":null,"abstract":"Mechanistic studies of biomolecular machines involved in intracellular protein degradation-such as the caseinolytic protease P, ATPases associated with diverse cellular activities (AAA+) motors, and the high-temperature requirement A family of enzymes-are of great interest as they are implicated in a host of human diseases. The function of these systems is dependent on both their fine-tuned three-dimensional structure and the conformational dynamics that modulate this structure. Their large sizes, inherent conformational plasticity, and oligomeric heterogeneity dictate that their mechanism of action cannot be deciphered by any one method. Synergistic application of methyl-transverse relaxation optimized spectroscopy (methyl-TROSY), nuclear magnetic resonance (NMR), and single-particle electron cryomicroscopy (cryo-EM) has uniquely positioned researchers to tackle the outstanding questions in this area of structural biology. Cryo-EM enables structural characterization and modeling of the large and conformationally heterogeneous complexes involved in protein degradation, while methyl-TROSY NMR enables monitoring structural transitions and conformational dynamics of these systems in response to various stimuli in solution at atomic resolution. This review highlights how combining these two approaches offers a distinct and powerful means to unravel allosteric pathways within complex, multipartite biomolecular machines.","PeriodicalId":8825,"journal":{"name":"Biochemical Journal","volume":"483 17","pages":""},"PeriodicalIF":4.3,"publicationDate":"2025-08-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12493191/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144940773","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Lipid nanoparticle-delivered intrabodies for inhibiting necroptosis and pyroptosis. 脂质纳米颗粒递送体内抑制坏死和焦亡。

IF 4.3 3区生物学

Biochemical Journal Pub Date : 2025-08-20 DOI: 10.1042/BCJ20253191

Veerasikku Gopal Deepagan, Xiuquan Ma, Farzaneh Bazregari, Jiyi Pang, Jan Schaefer, Joanne M Hildebrand, Ruby K Dempsey, Marcel Doerflinger, Christopher A Baldwin, Florian I Schmidt, James M Murphy, Ranja Salvamoser, James E Vince

{"title":"Lipid nanoparticle-delivered intrabodies for inhibiting necroptosis and pyroptosis.","authors":"Veerasikku Gopal Deepagan, Xiuquan Ma, Farzaneh Bazregari, Jiyi Pang, Jan Schaefer, Joanne M Hildebrand, Ruby K Dempsey, Marcel Doerflinger, Christopher A Baldwin, Florian I Schmidt, James M Murphy, Ranja Salvamoser, James E Vince","doi":"10.1042/BCJ20253191","DOIUrl":"10.1042/BCJ20253191","url":null,"abstract":"Intrabodies are intracellularly expressed high-affinity protein binders such as nanobodies and monobodies that offer an alternative approach to small molecules. However, the maturation of intrabody technology into new therapeutic modalities has been limited by the availability of a clinically relevant delivery system enabling sufficiently high levels of protein to be expressed in the cytosol. Here, we use lipid nanoparticle (LNP) systems based on clinically approved formulations for the efficient intracellular delivery of mRNAs encoding for intrabodies targeting mixed lineage kinase domain-like pseudokinase (MLKL) and apoptosis-associated speck-like protein containing a CARD (ASC), key mediators of the necrotic cell death modalities, necroptosis and pyroptosis, respectively. LNP delivery of intrabody mRNA resulted in robust protein expression, with an MLKL-binding intrabody preventing MLKL membrane translocation and protecting against necroptotic cell death. Similarly, LNP delivery of a bivalent intrabody targeting the inflammasome adaptor protein ASC protected against NLRP3 and AIM2 inflammasome-driven responses, including caspase-1 and IL-1β activation and gasdermin D-driven pyroptotic killing. These findings establish that LNPs harbouring anti-necrotic intrabody mRNAs allow for sufficient intracellular expression to neutralize necrotic cell death signalling and provide a general, clinically relevant, strategy for delivering therapeutic intrabodies into cells.","PeriodicalId":8825,"journal":{"name":"Biochemical Journal","volume":" ","pages":""},"PeriodicalIF":4.3,"publicationDate":"2025-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12493153/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144815746","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0